Application of CHAPS |CAS：75621-03-3|in protein extraction

CHAPS is an organic compound with molecular formula C32H58N2O7S and molecular weight 614.877.CHAPS is a kind of zwitterionic detergent, widely used in protein purification process. CHAPS can also be used in biochemical research to denature and split cells, extract RNA and DNA; In addition, CHAPS can also be used for the preparation of buffer solution.CN106478786A patent discloses a method for extracting effective intracellular protein of Desulfurization bacterium. CHAPS is used as amphoteric surfactant in the patent. The specific steps of the scheme are as follows:
     (1)Desulfurization bacteria activation culture, liquid fermentation culture, take the fermentation broth, centrifuge, remove the supernatant, and collect the lower sediment;
     (2)Take the precipitate in the middle and lower layers of step (1), add 1~2mL/g of cracking solution(The cracking liquid is the aqueous solution of urea, amphoteric surfactant and reducing protective agent, in which the amphoteric surfactant is CHAPS), stir it magnetically at 0~4 ℃, and put it into - 18~- 22 ℃ for 8~10 hours.
     (3)The frozen substance in step (2) is ultrasonically broken at 0~4 ℃, and then placed again at - 18~- 22 ℃ for 8~10 hours;
     (4)The frozen substance in step (2) is ultrasonically broken at 0~4 ℃, and then placed again at - 18~- 22 ℃ for 8~10 hours;
     (5)Take the supernatant protein solution obtained by centrifugation in step (4), add the organic mixed solution, fully mix and shake, and centrifuge;
     (6) After centrifuging the mixture in step (5), remove the supernatant, add organic solvent, fully mix and shake, and centrifuge the mixture;
     (7)Repeat the operation step (6) for 2-4 times, remove the supernatant from the centrifuged mixture, freeze-drying with lyophilizer, seal it, and store it at 0-4 ℃.
This patent realizes the separation and purification of effective intracellular protein of Desulfurization bacterium through centrifugation of fermentation broth, cell fragmentation, protein extraction and purification. It is an easy to operate, low cost, safe and reliable extraction method.

Reference
CN106478786A A Method of Extracting Effective Protein from Desulfurization Bacteria.